WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Histone H3 (tri methyl K4) Recombinant Rabbit mAb (S-1602-21)
Histone H3 (tri methyl K4) Recombinant Rabbit mAb (S-1602-21)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P68431 |
| Clone Number | S-1602-21 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ChIP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | HeLa, NIH/3T3, C6 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| Dot Blot | 1:1000 | |
| WB | 1:1000-1:5000 | Hu, Ms, Rt |
| IHC-P | 1:5000 | Hu, Ms, Rt |
| ChIP | 1:50 |
Background
Histone H3 (tri-methyl K4) is a chromatin-associated protein that has been modified by the addition of three methyl groups to the lysine residue at position 4. This post-translational modification is typically associated with active transcription, as it creates a binding site for chromatin remodelers and transcription factors, thereby promoting an open chromatin structure that facilitates gene expression.
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Western Blot
WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Dot Blot
Dot blot result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Lane 1: H3K4me3 modified peptide
Lane 2: H3K4 unmodified peptide
Lane 3: H3K4me1 modified peptide
Lane 4: H3K4me2 modified peptide
Primary antibody: Histone H3 (tri methyl K4) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human thyroid cancer. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human squamous cell carcinoma. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse colon. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat colon. Anti-Histone H3 (tri methyl K4) antibody was used at 1/5000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3 (tri methyl K4)
Recombinant Rabbit mAb (S-1602-21) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA) showed the enrichment of RPL30, GAPDH, MYOD1,
AFM, SAT-α and SAT-2 in Histone H3 (tri methyl K4) Recombinant Rabbit mAb (S-1602-21)-immunoprecipitated
sample.
